共表达 ORF67 和 ORF69 基因产物可重建卡波西肉瘤相关疱疹病毒核出芽复合物并形成核膜囊泡。
Reconstitution of the Kaposi's sarcoma-associated herpesvirus nuclear egress complex and formation of nuclear membrane vesicles by coexpression of ORF67 and ORF69 gene products.
机构信息
Viral Oncology Program, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Johns Hopkins University, School of Medicine, Baltimore, Maryland, USA.
出版信息
J Virol. 2012 Jan;86(1):594-8. doi: 10.1128/JVI.05988-11. Epub 2011 Oct 19.
Abstract
The Kaposi's sarcoma-associated herpesvirus nuclear egress complex is composed of two proteins, ORF67 and ORF69. In this study, we have recapitulated the KSHV complex by coexpression of these two proteins in insect cells using expression from recombinant baculoviruses. The proteins form a complex at the nuclear membrane as judged by live-cell analysis of protein fusions tagged with green fluorescent protein (GFP) and mCherry. Ultrastructural analysis of infected cells showed that ORF67 expression results in reduplication of the nuclear membrane. When the two proteins are expressed together, numerous virion-size nuclear membrane-derived vesicles were evident at the nuclear margins.
摘要
卡波氏肉瘤相关疱疹病毒核出芽复合物由两个蛋白，ORF67 和 ORF69 组成。在这项研究中，我们通过使用重组杆状病毒在昆虫细胞中表达这两种蛋白来重现 KSHV 复合物。通过用绿色荧光蛋白（GFP）和 mCherry 标记的蛋白融合的活细胞分析判断，这些蛋白在核膜上形成复合物。超微结构分析感染细胞显示，ORF67 的表达导致核膜的重复。当两种蛋白一起表达时，在核边缘可以明显看到许多病毒大小的核膜衍生小泡。
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